Construction Of Subtractive CDNA Library Of Feather Color Traits And Study On Differentially Expressed Genes In Mule Duck

In this experiment, the mule duck was chosen as the experimental material combined withthe method based on subtractive hybridization of the DSN mediated subtractive hybridizationin order to construct a subtracted cDNA llibrary of mule duck feather color related genes.Thepurpose of study would like to provide directions for the exploitation of the molecular mechanism offeather color related genes expression.In the study one subtractive hybridization library that related feather color development ofmule ducks were constructed in the experiment. The typical black-white Muscovy wasselected to conduct DSN-mediated subtractive hybridization in which the white hair follicletissue was considered as tester compared to black ones. During the experiment the reductionproducts was connected with a pUCm-T carrier through the blue-white screening.144positive clones were randomly selected in which about500-2000bp was selected by bacilliPCR and then sequenced. Finally a totall of64independent differentially expressed geneswere obtained in which the vector sequence and the redundant sequence were removed inanalysis by BLAST, and was found that21genes have homologous genes. By gene ontologyit showed that these21genes were related with cell communication and signal transduction,cytoskeleton and cell movem-ent, apoptosis, cell and body immune, transcriptional regulation.Another43with no homo-logous or new genes, along with while the functions of other geneshave not been reported yet.In order to identify the reliability about the difference of expression gene of SSH library,5genes from above constructed library was analyzed by RT-PCR. The white and blackfeather follicle skin cDNA that were acquired form3ten-weeks-old black and white muleduck was made as a template with the β-actin for control. The purpose was to detect how#26,#61,#160,#209and#240of these five genes in library about the white hair follicle andblack hair follicle express. The results were that the five genes expressed in the white hairfollicle were sign-ificantly higher than ones of black hair follicle(p<0.01). It was suggestedthat the cDNA library constructed in this experiment was of high reliability and good quality,which can be effectively enriched in differentially expressed genes of black and whiteplumage color.